Public EST Project for Soybean
Record of Deposit Gm-c1036
Soybean cDNA Library
Somatic embryos
The soybean cDNA library is being made publicly available by the following laboratory. To insure
that credit is given to the laboratory and scientists donating this valuable resource to the scientific
community, the information provided with the library must be maintained and included with any
transfer of biological materials or data derived from the library.
Library Source/Contributor
Dr. Lila O. Vodkin
University of Illinois at Urbana-Champaign
Department of Crop Sciences
384 Edward R. Madigan Laboratory
1201 W. Gregory Drive
Urbana, IL 61801-
Phone: 217-244-6147
Fax: 217-333-4777
e-mail: l-vodkin@uiuc.edu
Genotype/Cultivar
Jack
Vector Information
The cDNA library was prepared using the pSPORT 1 vector (Life Technologies) to
generate a plasmid library. The vector provides ampicillin resistance and allows blue-white color
selection for recombinant plasmids. The vector also contains unique Sal I and Not I sites needed
to clone cDNA directionally. The cloned cDNA fragment can be amplified by the polymerase
chain reaction using one of the following vector primer pair combinations: M13 universal and M13
reverse or T7 and SP6.
Library Description
The cDNA library was constructed from mRNA isolated from somatic embryos (age
ranging from 2 months to 9 months) cultured on MSD 20. The library was prepared using the Life
Technologies pSuperScript cDNA library construction kit. Complementary DNA was synthesized
from mRNA using a poly (dT) sequence with a Not I restrictions site. Sal I linkers adapters were
ligated to the blunt-ended cDNA fragments followed by Not I digestion. The cDNA fragments
were directionally cloned into the Not I-Sal I restriction site of the pSPORT 1 vector. The ligated
cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. This library was
constructed in the laboratory of Dr Lila Vodkin by Anu Khanna at the University of Illinois at
Urbana-Champaign.
e-mail: l-vodkin@uiuc.edu
Transformation efficiency: 10 6
Percent white colonies: 2%
Average insert size of white colonies: 1.23Kb
Disposition: These clones are being sent so the plates can be replicated. Send one plate back to
the originating laboratory and one plate to John Martin or Marco Marra at Washington University
for sequencing. All clone identifiers and shipping instructions are to be determined by Genome
Systems and Washington University.
NAME ASSIGNED BY GENOME SYSTEMS: Gm-c1036
Information for Genome Systems:
LABEL AND DATE ON TUBE SENT x NUMBER OF TUBES SENT:
cDNA (Somatic embryo) 2 tubes, 1/4/00
DILUTION INFORMATION: 100 l of the enclosed dilution of the library will give 2000 colonies.
Please plate on X-gal/IPTG plates and pick ONLY the white colonies. Initial tests show <10%
blue colonies.
PICKING INSTRUCTIONS: These cells are being sent so that colonies can be picked robotically
and entered into 384 well plates for preliminary analysis and eventually for high density filter
arrays. Please pick up to 5000 colonies. Send one set of plates back to my laboratory and keep
one set at Genome Systems.
Disposition: These clones are being sent so the plates can be replicated. Send one plate back to
the originating laboratory and one plate to John Martin or Marco Marra at Washington University
for sequencing. All clone identifiers and shipping instructions are to be determined by Genome
Systems and Washington University.